Adenosine Triphosphate Secondary article

Mildred Cohn, University of Pennsylvania, Philadelphia, Pennsylvania, USA Article Contents

. Introduction
Adenosine triphosphate consists of the purine adenine linked through its N9 to D-ribose via . Structure of ATP and its Metal Complexes
a b-N-glycosidic C(1’) linkage; the a-phosphoryl group is linked via an ester linkage to the . Energetics of ATP Hydrolysis
alcoholic O of ribose (C5’) and the b- and g-phosphoryl groups form anhydride bonds. The . ATP Synthase
chemical energy stored in the anhydride bonds is released upon hydrolysis at the g- or b- . Catalysis
phosphorus. . ATP in Metabolism
. ATP in Biosynthesis

Introduction . Regulation by ATP

. Energy Transduction by ATP Hydrolysis

Adenosine triphosphate (ATP), shown in Figure 1, is

ubiquitous in cells of living organisms. Of all the
biomolecules, its rate of turnover is the greatest; an average its negative charge, three or four negative charges at
person at rest produces and consumes about half his or her physiological pH. Not only is it consequently retained in
weight of ATP per day and the amount increases many fold the cell, the highly negatively charged phosphoanhydride
with strenuous activity. bonds are much less vulnerable to nucleophilic attack by
The concept of ATP as a ‘high energy’ compound, and hydroxide ion or water than, for example, carbonic
its central role in the storage of chemical energy that is anhydrides. Its negative charge also permits its binding
subsequently utilized for most essential biological pro- to metal ions and positively charged groups of proteins.
cesses requiring energy by cleavage of its phosphoanhy- The retention of thermodynamic instability coupled to
dride bonds, was first recognized by Lipmann in 1941. kinetic stability of the phosphoanhydride bonds in an
As indicated in Figure 2, ATP is formed mainly via aqueous medium is ideal for ATP’s role as a transmitter of
oxidative phosphorylation, photophosphorylation and free energy (Westheimer, 1987).
glycolysis. In anaerobic glycolysis, 2 moles of ATP are
produced per mole of glucose metabolized, but in oxidative
phosphorylation, 38 moles of ATP are produced for one
mole of glucose. The free energy of ATP hydrolysis to Structure of ATP and its Metal
adenosine diphosphate (ADP) and inorganic phosphate Complexes
(Pi) is utilized to maintain ion gradients and perform
mechanical work such as muscle contraction and flagella The structural features of ATP that remain invariant,
movement. The transfer of the g-phosphoryl group to whether in solution or crystals or whether liganded to
metabolites or proteins accompanied by the formation of
ADP is the most common mechanism of biological
Oxidative
regulation. In biosynthetic reactions such as fatty acid phosphorylation
activation, protein and RNA synthesis, the adenyl group of Glycolysis Photophosphorylation
ATP is transferred with the concomitant formation of
inorganic pyrophosphate (PPi), which is subsequently
hydrolysed enzymatically to two Pi. Thus with the free
ATP
energy available from the cleavage of two phosphoanhy-
dride bonds, the irreversibility of the synthesis is assured.
Of the inherent features of the structure of ATP that
make it suitable for its various functions, the primary one is
Biosynthesis Metabolic regulation
NH2
Dinucleotides Allosteric effector
N N Acy-CoA Covalent phosphorylation
N N Protein (metabolites, protein)
O– O– O–
γ β α
RNA
O– P O P O P O CH2 O Energy transduction
O O O
H
H H H
Mechanical work
OH OH (myosin, flagella, kinesin)
Active transport
Figure 1 Structure of ATP. The purine base, adenine, is linked to D-ribose Bioelectricity
by a b–N glycosidic bond. The 5’ position of ribose is linked by an ester bond Bioluminescence
to Pa, which in turn is linked by an anhydride bond to Pb and Pg of a
pyrophosphate moiety. Figure 2 The processes that synthesize and utilize ATP.

ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net 1

 Adenosine Triphosphate
metals or proteins, include ribose as d-ribose and the
linkage to adenine, which is always a b-N-glycosidic
linkage. In solution, ATP exists in two conformations
about the glycosidic C(1’)–N bond, syn if N3 of adenine lies
above the plane of the sugar group and anti if N3 points
away from the sugar group. At equilibrium, the anti
conformation predominates. The single conformation syn
or anti, which ATP will adopt when bound to a protein is
determined by intermolecular interactions such as hydro-
gen bonding and stacking effects with aromatic residues.
The relevant ATP species at the catalytic site of enzymes
is Mg2+ ATP or another divalent ion complex. The dimeric
species in crystals of the binary complex, Na2ATP.3H2O,
and of ternary complexes of MgATP stabilized with two
molecules of 2,2’-dipyridylamine have been solved by X-
ray diffraction (Cini et al., 1984). The relevance of these
dimeric structures to metal–ATP–protein complexes is
dubious, since the ATP or metal–ATP which binds to
protein is always monomeric. In solution, the MgATP
complex is a mixture of various species, but the situation is
simplified in the enzyme-bound MgATP since the protein-
binding domain restricts the permissible conformations
and stereochemistry to a single species. The structure of the
conformation of ATP in such complexes has been
investigated directly by X-ray crystallography and by
nuclear magnetic resonance (NMR) in solution and
indirectly with substitution inert complexes of ATP with
trivalent cobalt and chromium and with a- and b-thio-ATP
complexes with hard and soft divalent metal ions. It has
been found that the metal ion is always coordinated to the
g-phosphate and usually to the b-phosphate as well and
occasionally to the a-phosphate. The type of coordination
is not related to the specific bond cleaved in the reaction.
The most striking conclusion from an overwhelming
number of studies is that although there are a number of
recognizable conformations shared by bound MgATP,
often with functionally different proteins, there are also
different conformations found even in a group of enzymes
that catalyse the same type of reaction.
Energetics of ATP Hydrolysis
As shown in Figure 1, ATP has three phosphoryl groups; the
first, designated a, is bridged by the oxygen on C5’ of
adenosine forming an ester bond and is followed by two
phosphoryl groups, b and g, joined by phosphoanhydride
bonds. The a–b and b–g bonds are known as ‘high energy’
or ‘energy-rich’ bonds, defined as bonds with a high
negative free energy of hydrolysis. The free energy of
hydrolysis, DG0’, for the g-phosphoryl group to yield ADP
and Pi at pH 7.4 and 1 mmol L 2 1 Mg2+ is 2 8800
cal mol 2 1 and for the b- and g- phosphoryl groups to yield
AMP and PPi at pH 7.0 and excess Mg2+ is 2 7700
cal mol 2 1 (Jencks, 1976). The value of DG is dependent on
2 ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net
the concentration of the reactants, the ionic species (pH)
and divalent ion concentration; consequently the intracel-
lular value of DG for the hydrolysis of the g-phosphoryl
group under physiological conditions will vary as those
parameters vary. The highly exergonic hydrolysis of the
phosphoanhydride bonds of ATP makes possible the
coupling of these reactions to endergonic processes such as
synthetic reactions, ion transport and muscle contraction.
The large free energy change generated by hydrolysis of
ATP arises from the differences in the stability of ATP and
its hydrolysis products. The sources of the stability
differences are (1) the destabilization caused by the
electrostatic repulsion between the charged groups of the
phosphoanhydride relative to its hydrolysis products and
(2) the greater resonance stabilization in the hydrolysis
products than in the phosphoanhydride since in the latter,
unlike its hydrolysis products, the two phosphoryl groups
must compete for the p electrons of their bridging oxygen.
ATP Synthase
As indicated in Figure 2, oxidative phosphorylation is the
main pathway that leads to the synthesis of ATP from ADP
and Pi. The endergonic synthesis is coupled to the free
energy of the electrochemical proton gradient across the
mitochondrial membrane created by oxidation via the
electron transport system. The enzyme from mitochondria,
chloroplasts and bacteria is a complex proton-translocat-
ing transmembrane protein consisting of two separable
major substructures, a membrane-bound unit, F0, which is
responsible for proton translocation and a soluble unit, F1,
which catalyses the back reaction, ATP hydrolysis, and
some minor components. The arrangement and dimen-
sions of the components of mitochondrial ATP synthase
are shown in Figure 3. F1, which forms a flattened sphere
with its domain of alternating a and b subunits, is linked by
a stalk to F0. F1-ATPase consists of five subunits with the
stoichiometry, a3, b3, g, d, e. The subunits and stoichio-
metry of Escherichia coli F0 correspond to a, b2, and c9–12
components. The more complex mitochondrial F0 is
composed of nine different subunits, a, b, c, d, e, f, g, F6
and A6L. The stalk contains one copy each of the subunits
oligomycin-sensitivity-conferring protein (OSCP), d, F6
and b.
The three catalytic sites residing on the b subunit of F1
are asymmetric although all three a and b subunits have
identical amino acid sequences. ATPase activity requires
the g subunit in addition to a3b3. To explain the catalytic
behaviour of ATP synthase, Boyer (1993) suggested the
binding change mechanism in which energy from the
proton gradient is utilized for the release of a tightly bound
ATP and all three catalytic sites of F1 (ATP binding, ADP
binding, and empty) are highly cooperative. Thus the
binding of ADP and Pi at one site causes the release of ATP

α β α which requires both F1 and F0 remains to be elucidated.
β β 80 Å F1
α
δ ε
γ
50 Å Stalk
Stalk
50 Å F0
F0
Figure 3 A schematic representation of ATP synthase. The F1 component
(a3b3gde) is arranged in a flattened sphere of alternating a and b subunits.
Together with the g subunit the rotatory motor is formed with the g subunit
as the drive shaft. F1 is connected to F0, which is responsible for proton
translocation, by a stalk consisting of the subunits oligomycin-sensitivity-
conferring protein (OSCP), d, F6 and b.
at the adjacent site. It was further found that all three sites
were biochemically equivalent from experiments determin-
ing the distribution of 18O in isoptomers of ATP and Pi in
exchange reactions. To reconcile the fact that the single g
unit had to interact with three catalytic sites, Boyer
proposed an unusual and ingenious mechanism, namely
that the proton translocation in the F0 component drives a
rotation of the axially located g subunit, permitting
interaction sequentially with each catalytic site and thus
inducing conformational changes in the b subunits.
When the crystal structure of the mammoth (3440
residues) F1-ATPase from bovine heart mitochondria was
solved by Walker and his co-workers (Abrahams et al.,
1994), it confirmed Boyer’s unorthodox mechanism. The
structure resembled a rotatory motor with a hexagonal
ring of a and b subunit pairs surrounding a drive shaft, the
g subunit. The asymmetry of the a and b subunits is
immediately obvious and is due to conformational
differences; one catalytic site is occupied by a nonhydro-
lysable analogue of ATP, adenosine-5’-(b, g-imido) tripho-
sphate (AMP-PNP), the second by ADP and the third is
empty. The most dramatic demonstration of the rotatory
mechanism came from the elegant experiment of Noji (Noji
et al., 1997) in which a fluorescent actin filament was
attached to the g subunit of an a3b3g complex fixed to a
glass surface. When ATP was added, more than a 100
counterclockwise rotations of the actin filaments could be
observed visually under a microscope. It is the smallest
rotatory motor known.
Definitive knowledge of the structure of F0 is not as
satisfactory as for F1, nor is the detailed nature of the
interaction of the two components for the rotational
catalysis understood. The mechanism of the dissipation of
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Adenosine Triphosphate
the proton gradient accompanying the synthesis of ATP
Catalysis
The role of ATP in enzymatic reactions usually involves the
cleavage of the phosphoanhydride bonds; most frequently
the anhydride bond between O and Pg is cleaved.
Occasionally the intact molecule acts directly as an
allosteric effector as in aspartate transcarbamoylase and
phosphofructokinase catalysis. There is at least one
example, the formation of S-adenosylmethionine from
the reaction of methionine with MgATP, of the cleavage of
the C–O– | –Pa ester bond with the release of all three
phosphoryl groups. In all cleavage reactions of ATP, a
divalent metal ion is required, usually Mg2+. Enzymes
catalyse the cleavage of the phosphoanhydride bonds by
nucleophilic attack of an O or N from the substrate or
enzyme on either Pa, Pb or Pg. The largest group, cleavage
at Pg, includes the ATP hydrolases coupled to energy
transduction, the ubiquitous phosphotransferases (ki-
nases) and synthetases. Protein kinases, i.e. where protein
is the acceptor of the phosphoryl group, account for 2% of
the total products of the yeast genome. The second largest
group, the adenylyltransferases, which attack at Pa,
includes synthetases, polymerases and cyclase. The pyr-
ophosphotransferases, which attack at Pb, are rare, and
only seven such enzymatic reactions are known (Frick
et al., 1994).
Not only is the conformation of the enzyme-bound form
of MgATP independent of the site of bond cleavage, but
the mechanism of the catalytic reaction is similarly
independent. Three types of mechanisms occur, as
summarized in Table 1: (I) a single displacement, a direct
transfer from MgATP to an acceptor where both
substrates bind at adjacent sites; (II) a double displacement
with the formation of a covalent phospho- or adenylyl-E
intermediate and the release of ADP or PPi respectively,
followed by a second step with group transfer to an
acceptor; (III) activation by MgATP to form a phosphory-
lated or adenylylated product with the release of ADP or
PPi respectively, followed by a second step of ligation with
another substrate and the release of Pi or AMP. The first
two mechanisms can often be distinguished by kinetic
analysis but the stereochemical course of the reaction is an
unequivocal criterion for distinguishing them. Pa and Pb
can be made chiral by substitution of one of their oxygens
with S, 17O, or 18O and Pg by substitution of 2 O’s with S
and 17O or 18O.
Reactions of mechanism I proceed with inversion and of
mechanism II with retention of configuration. Some
phosphoryl transfer reactions proceed with inversion
(adenylate kinase) and some with retention (nucleoside
diphosphokinase); similarly for nucleotide transfer
3
 Adenosine Triphosphate
Table 1 Three mechanistic classes of ATP enzymes: nucleophilic attack at (A) Pγ , (B) Pβ and (C) Pα
I II
Simple transfer Covalent P-enzyme intermediate
(A) γ P: ATP + X → X-P + ADP (A) ATP + E → E-P + ADP
X = hydroxyl, carboxylate, amine E-P + X → X-P + E
phosphate, water X = hydroxyl nucleoside phos-
phate, water
(B) βP: ATP + X → X-PP + Pi (B) ATP + E → E-PP + AMP
X = hydroxyl E-PP + X → X-PP + E
X = hydroxyl
(C) αP: ATP + X → X-AMP + PPi (C) ATP + E → E-AMP + PPi
X = carboxylate, phosphate, sulfate, E-AMP + X → X-AMP + E
hydroxyl, water X = hydroxyl nucleoside phosphate
reactions, uridine diphosphate (UDP)-glucose pyropho-
sphorylase proceeds with inversion and galactose-P uridyl
transferase with retention.
The structure of the metal-ATP binding domain on
enzymes has been widely investigated by X-ray crystal-
lography and in solution by NMR. Several generalizations
have resulted:
1. A universal binding domain structure, i.e. a common
spatial protein fold and/or sequence motif, for ATP
does not exist.
2. Families of functionally unrelated enzymes may share
a common binding domain, the most common of
which resembles the Rossmann-dinucleotide fold,
alternating a helices and parallel b sheets, including
adenylate kinase, myosin, F1 ATPase, transport
proteins, enzymes involved in DNA recombination,
repair and replication and many others.
3. Enzymes closely related functionally may have more
than one binding motif; for example, aminoacyl-
tRNA synthetases are divided into two classes with
different binding motifs (Carter, 1993).
4. Some enzymes, for example pyruvate kinase, have a
completely unique binding domain.
ATP in Metabolism
Almost every metabolic pathway including carbohydrate,
lipid, amino acid and nucleotide metabolism involves ATP
directly as a substrate, or as a regulator. Its regulatory role
will be discussed in a subsequent section. The reaction
types in metabolism will be designated according to the
classification in Table 1. It was the dissection of the
glycolytic pathway, the conversion of glucose to two
pyruvates and the concomitant production of two ATP
4 ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net
III
Synthetases and ligases
(A) ATP + X + Y → X-Y + ADP + Pi
X = carboxylate
Y = amine, ammonia
(B) None
(C) ATP + X + Y → X-Y + PPi + AMP
X = hydroxyl, phosphate
Y = acyl, tRNA, CoA
molecules, into its ten enzymatically catalysed reactions,
that led to the discovery of ATP. The overall reaction is
given in reaction [IV].
Glucose + 2NAD+ + 2ADP + 2-
PI!2 pyruvate + 2NADH + 2ATP + 2H2O + 4H+
[IV]
In the first step, one ATP is consumed to form glucose 6-
phosphate and another ATP is consumed subsequently
producing fructose-1,6-bisphosphate. The loss of two
phosphoanhydride bonds is compensated for in later
reactions by the formation of four ATP, two from two
1,3-bisphosphoglycerates and two from two phosphoe-
nolpyruvates, catalysed by 3-phosphoglycerate and pyr-
uvate kinases, respectively. All ATP reactions of glycolysis
are type IA. The reactions of glycogen leading to glucose 6-
phosphate for entry into the glycolytic pathway do not
involve ATP as a substrate.
In the urea cycle, the mechanism most commonly used in
animals for getting rid of the nitrogen formed during the
catabolism of amino acids, three ATPs are used. The
overall urea cycle reaction is given in reaction [V].
Aspartate + 3ATP!urea + fumarate + 2AD
P + 2PI + AMP + PPi [V]
In the first step, carbamoyl phosphate is formed:
HCO32 + 2ATP + NH3!carbamoyl-P + 2ADP + Pi,
which includes two type IA reactions. In a subsequent step,
arginosuccinate, AMP and PPi are formed from citrulline,
ATP and aspartate with the intermediate formation of
citrullyl-AMP (type IIIC).
Prior to oxidation, fatty acids react with ATP in a type
IIIC reaction (reaction [VI]).
Fatty acid + ATP + CoA!Acyl-CoA + AMP + PPi
[VI]
with the intermediate formation of acyladenylate. Pro-
ducts of carbohydrate, amino acid and lipid metabolism all

feed into pyruvate, which is metabolized via the citric acid
cycle. One of its eight enzymes involves ATP, succinyl-
CoA synthetase of plants and bacteria (the mammalian
enzyme utilizes guanosine triphosphate (GTP)). Overall it
falls into the type IIIA class: succinate + ATP + -
CoA!succinyl-CoA + ADP + Pi, but there are three
steps, the initial formation of E-P (type IIA) followed by
transfer of the phosphoryl group to succinate and P-
succinate, which reacts with CoA. A unique reaction is
encountered in the synthesis of adenosine-3’,5’-cyclic
monophosphate (cAMP) from ATP through the attack
on the a-P intramolecularly by the 3’-OH of ATP catalysed
by adenylate cyclase.
ATP in Biosynthesis
The large negative free energy of hydrolysis of the
phosphoanhydride bonds of ATP is used to drive
endergonic syntheses of both small molecules and macro-
molecules. Among the small molecules are nucleotides,
coenzymes and cholesterol. The purine nucleotides AMP
and guanosine monophosphate (GMP) are derived from
inosine monophosphate (IMP). Of the eleven reactions
leading to IMP, five involve ATP. In the first reaction, the
phosphoanhydride bond of 5-phosphoribosyl-a-pyropho-
sphate is formed from ribose 5-phosphate in the rare
nucleophilic attack on the b-P of ATP (type IB). Four
subsequent steps 3, 5, 6, and 8 of this pathway are
synthetases (type IIIA). After IMP formed in this pathway
reacts with NAD+, the xanthosine monophosphate
formed reacts with ATP and glutamine to yield GMP.
The nucleoside monophosphates are converted to dipho-
sphates (NDP) by a dismutation reaction with ATP
yielding two NDPs (type IA). Subsequent reaction of
NDP with ATP forms a phosphoanhydride bond, cata-
lysed by nucleoside diphosphokinase, producing nucleo-
side triphosphate (NTP) and ADP. In reactions that utilize
triphosphates other than ATP, the requisite NTP is
supplied by this reaction. Pyrimidine monophosphates,
uridine and cytidine monophosphates (UMP and CMP)
are synthesized by a different pathway consisting of six
reactions. In animals the first three steps are catalysed by a
multifunctional enzyme on a single polypeptide chain.
Only the first step in UMP synthesis involves ATP, the
formation of carbamoyl phosphate in the reaction of 2ATP
with HCO32 and glutamine in a type IIIA reaction. UMP is
converted to uridine triphosphate (UTP) by two sequential
kinase steps as for the purines. Cytidine triphosphate
(CTP) is generated from UTP by amination.
The coenzyme flavin–adenine dinucleotide (FAD) is
synthesized in two steps. Riboflavin reacts with ATP in a
kinase reaction (type IA) to form riboflavin mononucleo-
tide (FMN) and ADP. The phosphoryl group of FMN
then accepts the adenylyl group of an ATP to yield flavin
ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net
Adenosine Triphosphate
adenine dinucleotide in a type IC reaction. An analogous
ATP reaction with nicotinamide mononucleotide gener-
ates the coenzyme nicotinamide adenine dinucleotide.
Coenzyme A (CoA) is synthesized from pantothenate in
five steps, of which four have ATP as a substrate. The first
step, the phosphorylation of pantothenate (type IA), is
followed by reaction with ATP and cysteine in a synthetase
reaction (type IIIA) producing 4’-phosphopantothenoyl
cysteine. After decarboxylation, the 4’-phosphopan-
tetheine reacts with ATP, which transfers its adenylyl
group (type IC) to form a phosphoanhydride bond in
dephospho-CoA. In the last step, ATP donates its g-
phosphoryl group to the O on the 2’ carbon to form CoA
with its three phosphoryl groups.
Cholesterol, a precursor of steroid hormones and a
component of cell membranes, is synthesized by a complex
series of reactions from acetyl CoA with the intermediate
formation of isoprene units, squalene and lanosterol. Only
three reactions in the initial phase of the synthesis leading
to isopentenyl pyrophosphate utilize ATP. The first step is
the production of hydroxymethylglutaryl-CoA (HMG-
CoA) from acetyl CoA followed by reduction to mevalo-
nate and the expulsion of CoA. The mevalonate is then
phosphorylated by ATP in its 5 position in a kinase
reaction (type IA), and the product is further phosphory-
lated to form a pyrophosphate. In a type IIIA reaction
mevalonatepyrophosphate is decarboxylated yielding iso-
pentylpyrophosphate + carbon dioxide + ADP + Pi. The
examples of biosynthesis discussed above illustrate that all
types of reaction listed in Table 1 may be involved.
The synthesis of macromolecules is often complex but
the initial step always involves ATP and/or other NTPs. In
RNA synthesis all four NTPs (for DNA deoxy-NTPs) are
required, each contributing a nucleotidyl group to form a
3’-5’ diester linkage and concomitantly PPi, as represented
by the type IC (reaction [VII]).
(RNA)n units + NTP!(RNA)n + 1 units + PPi [VII]
Amino acid activation, the first step in protein synthesis, is
a type IIIC reaction with an aminoacyladenylate inter-
mediate (reaction [VIII]).
Amino acid + ATP!aminoacyladenylate + PPi
[VIII]
Aminoacyladenylate + t-RNA!aminoacyl t-
RNA + AMP
ATP also functions in later steps of protein synthesis
including some pathways of protein folding involving
chaperonins such as Gro EL of E. coli. In the synthesis of a
typical diacylglycerophospholipid, phosphocholine at-
tacks the a-P of CTP to form CDP-choline and PPi (type
IC). UTP is cleaved at its a-P in the first step of glycogen
synthesis to convert glucose 1-phosphate to UDP-glucose
and PPi. All these syntheses involve nucleotidyl transfer
5
 Adenosine Triphosphate

with the release of PPi, which is hydrolysed to Pi ensuring
that the syntheses are driven to completion.
Regulation by ATP
ATP may regulate activity in three ways: (1) direct
allosteric effector; (2) covalent phosphorylation of serine,
threonine, tyrosine histidine or lysine residues of proteins
by protein kinases; (3) protein conformational changes
between its ATP and ADP complexes subsequent to
hydrolysis corresponds to active and inactive forms. An
example of ATP as a direct allosteric effector is the increase
of activity, upon binding of ATP of aspartate transcarba-
moylase, which catalyses the first step in pyrimidine
synthesis. A single phosphorylation of a serine residue of
pyruvate dehydrogenase by ATP controls its activity; the
phospho-enzyme is inactive and the dephospho-enzyme
regains activity. The number of regulatory mechanisms
which depend on protein phosphorylation is legion.
Control of glycogen metabolism by glycogen phosphor-
ylase, which effects its breakdown, and glycogen synthase,
which effects its synthesis, illustrates regulation by both
allosterism and a cascade of covalent phosphorylations. In
muscle, phosphorylase is allosterically activated by AMP
and inhibited by ATP. The kinase, which converts the
enzyme to an active form by phosphorylating its serine 14,
is itself activated by a cAMP-dependent protein kinase.
For glycogen synthase, the dephosphorylated form is the
active one. The phosphorylated form is strongly inhibited
by ATP. A more complex cascade controls the activity of
the glycogen synthase including a cAMP-dependent
protein kinase. The control of the breakdown and synthesis
are coordinated through their common controls, phos-
phorylation which activates phosphorylase and inactivates
the synthase; the two cascades are also linked. cAMP
synthesis, in turn, is controlled by a cascade of signal
transduction initiated by the binding of hormones to their
receptors which activate G proteins (GTPases whose
activity is switched on and off by displacing bound GTP
with GDP), and in the next step, G proteins activate
adenylate cyclase. For the complex enzyme nitrogenase,
analogous to G proteins, ATP and ADP act as an activity
switch (Howard and Rees, 1994).
Energy Transduction by ATP Hydrolysis
The processes that utilize ATPases to convert the stored
chemical energy in ATP to other forms of energy are shown
in Figure 2. To maintain active transport across a membrane
against a concentration gradient, hydrolysis of ATP may
be the direct or indirect source of energy. In the Na+ pump,
Na+-K+ATPase acts directly by phosphorylating an
aspartate residue of the enzyme only in the presence of
Na+ (type IA) and hydrolysing E-P only in the presence of
K+. A similar mechanism obtains for transporting Ca2+
to create an electrochemical potential gradient across a
membrane. These processes are the inverse of ATP
synthase where the energy from the dissipation of the
proton gradient is coupled to the synthesis of ATP. Since
the transport of glucose is supported by a Na+ gradient
which is generated by ATP hydrolysis, it is indirectly ATP-
driven.
ATP hydrolysis is the source of power for mechanical
motion in muscle contraction and the beating of flagella
and cilia. Contraction of vertebrate striated muscle is the
most thoroughly investigated motion. The accepted model
for the contractile force is that interpenetrating alternate
sets of thick and thin filaments slide past each other, a
linear motor rather than the rotary motor of ATP synthase.
The force originates from hydrolysis of ATP. The thick
filaments are composed of several hundred myosin
molecules, a large protein made up of heavy chains,
essential and regulatory light chains. A subfragment (S1),
which retains myosin’s ATPase activity, may be isolated
after two sequential proteolytic steps on myosin. Actin is
the primary component of the thin filaments. The thick and
thin filaments associate by cross bridges in the S1 domain
of myosin. The myosin-catalysed hydrolysis rate of ATP is
accelerated 20-fold by actin. Taylor and Lymn (1972)
formulated the following scheme: ATP binds to the S1
domain of actomyosin, which releases actin and generates
the complex, myosin–ADP–Pi. With the release of Pi from
the complex, the S1 head binds strongly to actin, powering
the sliding motion, followed by the release of ADP, and the
cycle begins again. Other types of motility are also
mediated by ATP-driven motors. As indicated in Figure 2,
the generation of bioluminescence of the firefly in the
luciferin–luciferase system is also coupled to the hydrolysis
of ATP to AMP and PPi.
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Alberts B, Bray D, Lewis I et al. (1994) Molecular Biology of the Cell, 3rd
edn, Chapters 5 and 15. New York: Garland Publishing.
ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net
Adenosine Triphosphate
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Ernster L (ed.) (1984) Bioenergetics. Amsterdam: Elsevier Science
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